文献：使用光镊雕刻和融合仿生囊泡网络

链接：https://www.nature.com/articles/s41467-018-04282-w

作者：吉多·博洛涅西马克·S·弗里丁阿里·萨利希-雷哈尼内森·E·巴洛，尼古拉斯·J·布鲁克斯，奥斯卡·塞斯尤瓦尔·埃拉尼

节选：

囊泡融合和囊泡微反应器

为了制备功能化的金纳米粒子囊泡，我们首先在含有2 wt.% 16:0 生物素帽聚乙烯 (Biotinyl Cap PE) 的0.5 M蔗糖溶液中电铸POPC囊泡。当存在荧光脂质时，其含量为1 wt.%。在钙黄绿素稀释和蛋白质表达实验中，囊泡通过乳液相转移（见上文）形成，浓度为0.5 M蔗糖/葡萄糖密度梯度。接下来，将150 nm链霉亲和素包被的金纳米粒子（Nanopartz，美国科罗拉多州；产品C11-150-TS-50；2.5 mg ml −1）以1:9的比例添加到囊泡中，将样品涡旋30分钟以促进结合。VIM的形成方法与之前相同，但外部溶液中的NaCl含量为0.25 M。通过在VIM界面上聚焦并施加150 mW（在阱处）或更高功率的激光来实现囊泡的融合。为了避免意外融合，使用小于100 mW的激光（在阱处）对囊泡进行空间操控（补充说明 13）。我们还使用了Aurora-DSG纳米粒子（AuNP标记的脂质；1-2 nm；Avanti），当其以2 wt.%的浓度嵌入膜中时，不会发生融合。

Vesicle fusion and vesicle microreactors

To prepare functionalised AuNP vesicles, we first electroformed POPC vesicles in 0.5 M sucrose solution with 2 wt.% 16:0 Biotinyl Cap PE. When fluorescent lipids were present, these were at 1 wt.%. In the calcein dilution and protein expression experiments, vesicles were formed via emulsion phase transfer (see above), with 0.5 M sucrose/glucose density gradient. Next, 150 nm streptavidin-coated AuNPs (Nanopartz, CO, USA; product C11-150-TS-50; 2.5 mg ml−1) were added to the vesicles 1:9, with the sample vortexed for 30 min to drive conjugation. VIMs were formed as before, but with 0.25 M NaCl in the external solution. Vesicles were fused by focusing and applying a laser of 150 mW (at trap) or greater on the VIM interface. Vesicles were manipulated in space using a <100 mW laser (at trap) to avoid unintentional fusion (Supplementary Note 13). We also used Aurora-DSG Nanoparticles (AuNP-labelled lipids; 1–2 nm; Avanti), which did not result in fusion when embedded in the membrane at 2 wt.%.

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